Recombinant human cc10 and compositions thereof for use in the treatment of nasal rhinitis

ABSTRACT

The present invention relates generally to the use of recombinant human CC10 (rhCC10), also known as recombinant human uteroglobin, for use as a therapeutic in the treatment of nasal rhinitis, nasal sinusitis, chronic rhinosinusitis, and nasal polyposis. More particularly, the invention provides methods, including broadly the critical dosage ranges of rhCC10 and intranasal route of administration, which may be administered to safely and effectively treat the aforementioned conditions. The invention further provides a composition useful in administering rhCC10 to humans.

CROSS-REFERENCE TO RELATED APPLICATIONS

This application claims benefit of and priority to, U.S. ProvisionalPatent Application 61/052,861, filed May 13, 2008, the disclosure ofwhich is hereby incorporated by reference in its entirety.

FIELD OF THE INVENTION

The present invention relates to methods of reducing airflow obstructionin the nasal passages, clearing a sinus infection, and reducing sinuspain in a patient. More specifically the present invention relates tomethods of treating nasal rhinitis, sinusitis and nasal polyposis inpatients and compositions useful for the same. Yet more specifically,the present invention relates to methods of treating the above usingintranasally-administered recombinant human CC10 and compositionsthereof useful for the same.

BACKGROUND

Clara Cell “10 kDa” protein (CC10) or uteroglobin (UG) is a small,homodimeric secretory protein produced by several mucosal epithelia andother organs of epithelial origin (Mukherjee, 1999). CC10 consists oftwo identical subunits of 70 amino acid residues, each with the “fourhelical bundle” secondary structure motif, joined in antiparallelorientation by two disulfide bonds between Cys 3 and 69′, 3′ and 69(Matthews, 1994; Morize, 1997). The homodimer containing two disulfidebonds appears to be its primary, extracellular active form. In humans,the lung is the main site of CC10 production, while several other organssynthesize smaller amounts of mRNA encoding this protein (Singh, 1987;Sandmoller, 1994). CC10 is an anti-inflammatory and immunomodulatoryprotein that has been characterized with respect to various interactionswith other proteins, receptors and cell types (reviewed in Mukherjee,2007, Mukherjee, 1999, and Pilon, 2000). Lower levels of CC10 protein ormRNA have been found in various tissue and fluid samples for a number ofclinical conditions characterized by some degree of inflammationincluding asthma (Lensmar, 2000; Shijubo, 1999; Van Vyve, 1995),pneumonia (Nomori, 1995), bronchiolitis obliterans (Nord, 2002),sarcoidosis (Shijubo, 2000), and in patients suffering from chronicrhinitis with recurrent sinusitis and nasal polyposis (Liu, 2004).Pulmonary epithelial cells, the body's primary source for endogenousCC10, are often adversely affected in these conditions, depleted or evenablated (Shijubo, 1999). Indeed, CC10 appears to be an autocrine and/orendocrine required for development of specific sets of non-ciliatedrespiratory epithelial cells and associated structures (Castro, 2000).Thus, it is still not known whether CC10 deficiency is a cause or aneffect of the inflammation and/or the condition.

The obstruction of airflow in the nasal passages, as well as sinus painand pressure, are known to be causes of significant morbidity in humanssuffering from allergic rhinitis, non-allergic rhinitis, sinusitis, andnasal polyposis. Nasal rhinitis is an inflammation of the nasal passagesand sinuses in the nasopharygeal cavity. There are two types ofrhinitis, allergic and non-allergic. Non-allergic rhinitis is due toviral, bacterial, or other infection, to exposure to inhaled chemicalsor other irritants, or may be idiopathic, while allergic rhinitis is dueto exposure to inhaled allergens. Allergic rhinitis may be seasonal,such as allergy to tree or grass pollen; or it may be perennial, such asallergy to dust mites and common molds. Rhinitis ranges in severity frommild seasonal discomfort due to itching, sneezing, and nasal dischargefor a few hours, days or weeks, to painful and debilitating chronicsinus inflammation that is often associated with recurrent bacterialinfection. Chronic sinus inflammation in the presence of bacterialinfection is sometimes referred to as chronic rhinosinusitis (“CRS”).CRS, leads to irreversible remodeling and scarring of airway epitheliaand sinus tissue. These permanent changes to the nasal tissues result ina vicious cycle in which decreased ability to fight infection, bothviral and bacterial, as well as a decreased ability to clear inhaledallergens and irritants, lead to even more exaggerated inflammatoryresponses, further exacerbating remodeling and fibrosis, and more severeor persistent infections. Theoretically, inflammation is reversible inthe absence of infection, and should disappear as soon as the irritant,pathogen, or allergen is cleared from the local tissue. Therefore, thetransition from seasonal or mild rhinitis to CRS can be attributedlargely to chronic exposure to perennial allergens and/or recurrentbacterial infection of the inflamed nasal and sinus tissue, leaving theinfection to persist even after the original rhinitis stimuli (allergenor irritant) is long gone. Indeed, patients with perennial allergiesresulting in chronic rhinitis often experience recurrent bacterialinfection (sinusitis) as the inflammatory response transitions from anallergen-stimulated response to an infection-stimulated response. Theseare the patients with severe persistent rhinosinusitis CRS disease andthe highest morbidity. Chronic rhinitis, whether allergic ornon-allergic, results in excess mucus production in, and swelling of,nasal passages that impairs breathing, disrupts sleep, and predisposesto repeated bacterial sinus infections. The sinus pain and pressurecauses significant morbidity in this disease. Bacterial infection,whether acute or chronic, exacerbates these symptoms. In the most severecases, nasal polyps grow in the nasal airways and slowly obstruct them.These polyps are non-malignant outgrowths of sinus tissue which can onlybe removed by sinus stripping surgeries. A patient with nasal polyps mayundergo sinus stripping periodically since the polyps grow back aftereach removal.

However, determining whether rhCC10 can alleviate inflammation, and atwhat dosage, in patients suffering from nasal rhinitis, especiallychronic rhinitis and rhinosinusitis, with or without nasal polyposis andin patients suffering from chronic or recurrent bacterial sinusinfection has remained elusive. In fact, as shown below, recent workindicates that at dosages known and commonly used, rhCC10 isineffective:

In a recent Phase II clinical study to evaluation the efficacy ofintranasal rhCC10 to suppress nasal inflammation and rhinitis due toseasonal allergy, rhCC10 treatment resulted in a significant worseningof symptoms in one of six efficacy outcome measures compared to placebo(Widegren, et al., 2009). The remaining five efficacy outcome measuresshowed no difference between rhCC10 and placebo, although all trended infavor of placebo. RhCC10 was inferior to placebo in improving(increasing) peak nasal inspiratory flow and in mitigating rhinorreacaused by administration of aero-allergens. Table 1 shows comparativeoutcomes of patients while receiving rhCC10 versus outcomes in the samepatients while receiving placebo, as measured during the last three days(days 5-7) of each treatment period.

TABLE 1 P-values for statistical differences between rhCC10 and placeboin five clinical outcome measures. Efficacy Variable rhCC10 PlaceboP-value Morning TNSS 1.64 (0.21) 1.50 (0.25) .57 Morning PNIF 136 (7)136 (8) .78 Evening TNSS 1.37 (0.28) 1.54 (0.27) .53 Evening PNIF 145(8) 147 (8) .93 TNSS-10 min 5.67 (0.27) 5.17 (0.32) .09 after challengePNIF-10 min after 93 (6) 102 (7)  .04* challenge *A p-value of <0.05(less than 0.05) is considered to be a significant difference. P-valueshigher than 0.05 are not considered to be statistically significant.TNSS: Total nasal symptoms score PNIF: Peak nasal inspiratory flow

This proof-of-concept study failed to demonstrate the overall efficacyof rhCC10 given once daily for seven days in this nasal allergenchallenge model of seasonal allergic rhinitis. RhCC10, given 1.1 mg in200 μL per day intranasally, did not favorably affect allergen-inducedmorning, post challenge or evening symptoms compared with placebo. Ahigher PNIF reflects greater airflow and a lower PNIF indicatesrestricted airflow. Morning as well as evening PNIF were unaffected byrhCC10, however, post challenge PNIF was modestly reduced by rhCC10treatment compared to placebo, which did reach statistical significance.Symptom-scores and PNIF-levels reached in the placebo arm were verysimilar to those recorded historically in this model. Likewise, markersof inflammation in nasal lavage fluids, including levels of eosinophilcationic protein, myeloperoxidase, and alpha2-macroglobulin, and rhCC10did not mediate any reduction in these markers compared to placebo. Inthis model, it has been demonstrated that corticosteroids inhibitmorning, post-challenge as well as evening symptoms and these markers ofinflammation in nasal lavages (Ahlstrom-Emanuelsson et al., 2002 & 2007)whereas anti-histamines reduce post-challenge symptoms only (Korsgren etal. 2007). Therefore, using this dose, dosing regimen, volume and spraymethod of intranasal administration, rhCC10 did not demonstrateanti-allergy, anti-inflammatory effects in all six clinical outcomemeasures or in all three inflammatory markers in nasal lavages.

Currently, most nasal rhinitis and rhinosinusitis are treated withvarious over-the-counter and prescription medications such asanti-histamines, decongestants, non-steroidal anti-inflammatory agents(“NSAIDS”) and various non-pharmacologic nasal sprays and irrigationsolutions. Chromium nasal solutions, oral anti-histamines andleukotriene receptor antagonists treat symptoms but provide only a fewhours of relief. Nasal oxymetazoline solutions are very effective atopening nasal passages but overuse results in a “rebound effect” andrapid loss of efficacy with worsening of symptoms. Side effects forthese types of drugs include sore throat, dehydration of nasal tissues,and constipation, among others.

Furthermore, sinusitis is typically treated with oral antibiotics.Antibiotics range in side effects from mild to severe and can includeconstipation and other digestive problems, headache, dizziness, rashes,liver, kidney and bladder toxicity, muscle and joint pain, etc.Antibiotics can also cause hypersensitivity reactions, particularly inpatients with recurrent sinusitis who have to take antibioticsrepeatedly and eventually become allergic to them. Hypersensitivityreactions to antibiotics may occur without warning or previous signs ofallergy and may be suddenly lethal.

For severe and/or chronic rhinosinusitis disease, physicians currentlyprescribe nasal corticosteroids, which reduce inflammation but oftenlose efficacy after a few weeks or months of continuous therapy. Oralcorticosteroids are also efficacious but have many undesirable sideeffects when used for long periods of time. For example, in adults,cardiovascular complications, including hypertension and stroke, aremajor side effects of corticosteroid use. In children, corticosteroidsimpair normal growth and development. In all patients, corticosteroidslower the patient's immune function and leave them susceptible toinfection of all types (bacterial, viral, fungal, etc.). Thus, safety isa major consideration in the choice of drugs and drug combinations usedto treat, prevent or cure nasal rhinitis, especially chronicrhinosinusitis, nasal polyposis, chronic or recurrent bacterial sinusinfection, their associated morbidities and other similar conditions.

There are several formulations, devices, and methods by which drugs maybe administered intranasally to treat rhinitis, sinusitis, andrhinosinusitis. One method for administering local intranasal doses ofdrugs to the nasal passages and sinuses is the use of liquid drugformulations in spray bottle or spray pump devices that are converted toaerosols by being forced through a small aperture and sprayed into theanterior portion of the nasal cavity through each nostril.

Particle sizes generated by the aforementioned devices are in the 5-10micron range, which maximizes delivery and local deposition of the drugin the nasal mucosa lining the nasopharygeal cavity. The nasal mucosa iscomprised of a normally thin layer of mucus that overlays the wetepithelium in the nasal passages and sinuses of the nasopharyngealcavity. Most 5-10 micron particles sprayed into the nostril will impactthe non-ciliated epithelium in the anterior portion of thenasopharyngeal cavity. Once deposited at the site of impaction in thenasal mucosa, drugs may distribute throughout the mucosa and be clearedat various rates through the action of cilia and ciliated epithelialcells located in the posterior two thirds of the nasopharyngeal cavitythat push towards the pharynx where the drug and mucus are swallowed.The local action of drugs deposited in the nasal cavity depends upon theparticle size delivered, the formulation, and the rate of clearance.These factors affect the efficiency of local delivery and the length oftime that the nasopharyngeal mucosa and epithelia are exposed to thedrug before it is cleared.

The local action of intranasally administered drugs also depends uponthe condition of the nasal mucosa and tissues at the time of delivery.For example, when the nasal passages are blocked by thick mucus, localdelivery of drugs is very difficult, if not impossible.

Therefore, it is a significant challenge to find an agent, and a correctdosage for that agent, which alleviates airway obstruction, sinus painand discomfort for a pro-longed period of time without serious sideeffects. There is therefore a need for new, more effective orlonger-lasting agents and formulations thereof and administration anddosage regimens thereof, particularly in patients with chronic disease.

OBJECTS OF THE INVENTION

The foregoing provides a non-exclusive list of the objectives achievedby the present invention:

It is a primary object of the invention to treat, cure or prevent nasalrhinitis, sinusitis, especially chronic rhinitis and rhinosinusitis,with or without nasal polyposis, in patients using rhCC10.

It is a further object of the invention to alleviate the pain and sinuspressure associated with chronic rhinitis, sinusitis, and rhinosinusitisusing rhCC10.

It is a further object of the invention to enable patients with nasalrhinitis or sinusitis to achieve a better quality of sleep using rhCC10.

It is a further object of the invention to administer the rhCC10 topatients by intranasal instillation, nasal lavage or as an intranasalaerosol involving the use of a spray device.

It is a further object of the invention to provide a safe,well-tolerated and effective dosage range of rhCC10 which accomplishesthe above objectives and does not significantly suppress the immuneresponse or increase the frequency or severity of adverse events.

It is a further object of the invention to provide a drug-devicecombination by which rhCC10 can be effectively administered to thenasopharygeal cavity as an aerosol, as a gel, or as a liquid.

It is a further object of the invention to provide a formulation ofrhCC10 in specific pharmacologically-acceptable nasal excipients forapplication in a gel or cream for local administration and prolongedrelease, using a single use swab applicator.

It is a further object of the invention to provide a formulation ofrhCC10 in specific pharmacologically-acceptable nasal excipients forapplication as an aerosol with particles sizes in the 5-10 micron rangefor local administration and deposition, using a spray pump or squeezebottle.

It is a further object of the invention to provide a formulation ofrhCC10 in specific pharmacologically-acceptable nasal excipients forapplication as an aerosol with particles sizes in the 5-10 micron rangefor local administration and deposition, using a multi-use spray pumpdispenser, metered dose inhaler (MDI), or squeeze bottle device.

It is a further object of the invention to provide a formulation ofrhCC10 in specific pharmacologically-acceptable nasal excipients forapplication as an aerosol with particles sizes in the 1-5 micron rangefor pulmonary administration and deposition, using a single or multi-usespray pump dispenser, metered dose inhaler (MDI), or squeeze bottledevice.

It is a further object of the invention to provide a formulation ofrhCC10 in specific pharmacologically-acceptable nasal excipients forapplication as an instillation in a single or multi-dose syringe device.

It is a further object of the invention to provide a formulation ofrhCC10 in specific pharmacologically-acceptable nasal excipients forapplication as an instillation in a nasal lavage solution using a “netipot” or similar gravity flow lavage device.

It is a further object of the invention to provide rhCC10 itself as apharmacologically-acceptable nasal excipient for the alleviation ofpain, irritation, and discomfort caused by local inflammatory responsesat the site of administration of other drugs in the nasopharyngealcavity.

It is a further object of the invention to provide rhCC10 itself as apharmacologically-acceptable nasal excipient to enhance thebioavailability of other drugs administered to or via the nasopharyngealcavity.

It is a further object of the invention to provide rhCC10 as an activeingredient in combination formulations with other drugs for intranasaladministration.

The term “pharmacologically-acceptable” is intended to characterize aformulation or combination of excipients that cause no deleteriouseffects or cause deleterious effects that are known and are, or can be,accepted by regulatory authorities.

SUMMARY OF THE INVENTION

These and other objects, features and advantages are achieved byadministering rhCC10 in a dosage range given at appropriate intervals,or in one dose, to treat, cure or prevent nasal rhinitis, sinusitis,rhinosinusitis, and CRS, with or without nasal polyposis. Furthermore,for chronic rhinitis patients rhCC10 offers an even greater benefit intreating, curing or prevention of chronic rhinitis when given in adosage range at appropriate intervals, or in one dose. Thus, it has nowbeen surprisingly found that rhCC10, which was thought to be ineffectivein curing, treating or preventing nasal inflammation, rhinitis, nasalrhinitis, chronic rhinitis, sinusitis and rhinosinusitis, is in facteffective when used in accordance with the invention herein.

These and other objects, features and advantages are also achieved byadministering rhCC10 in a dosage range given at appropriate intervals orin one dose where a patient shows one or more of the following: sinuspain and pressure, inability to sleep due to sinus discomfort, chronicrhinitis, rhinosinusitis, and growth or regrowth of nasal polyps.

These and other objects, features and advantages are also achieved byadministering rhCC10 such that it does not inhibit platelet aggregation,suppress the immune response, such as in common cold or flu, or increasethe frequency or severity of any adverse event.

In certain aspects of the invention, rhCC10 is administered intranasallyin a single dose divided about equally between each nostril in a rangeof 1.5 micrograms to 1.1 milligrams per day, or in multiple doses whichtaken together achieve this dosage range on a daily basis to treat, cureor prevent severe nasal rhinitis, nasal sinusitis, especially chronicrhinosinusitis, and/or nasal polyposis. In another aspect, an intranasalrhCC10 dose or doses divided about equally between each nostril in arange of 1.5 micrograms to 1.1 milligrams per day can be repeated atappropriate intervals to treat, cure or prevent severe nasal rhinitis,nasal sinusitis, chronic rhinitis with recurrent sinusitis, especiallyrhinosinusitis, and/or nasal polyposis. In yet another aspect of theinvention, rhCC10 is administered intranasally on a daily basisconsecutively for seven days, ten days, 14 days, or 21 days.

In yet another aspect of the invention, rhCC10 is administered threetimes per day, at approximately eight hour intervals in intranasal dosesdivided about equally between each nostril in a range of 0.5 to 370micrograms per day. In yet another aspect of the invention, rhCC10 isadministered two times per day, at approximately twelve hour intervalsin intranasal doses divided about equally between each nostril in arange of 0.75 to 650 micrograms per day.

In yet another aspect of the invention, rhCC10 is administered in atapered fashion, beginning with three times per day, at approximatelyeight hour intervals in intranasal doses divided about equally betweeneach nostril in a range of 0.5 to 370 micrograms per day for three days,followed by two times per day, at approximately twelve hour intervals inintranasal doses divided about equally between each nostril in a rangeof 0.5 to 370 micrograms per day, followed by one time per day inintranasal doses divided about equally between each nostril in a rangeof 0.5 to 370 micrograms per day. In yet other aspect of the invention,rhCC10 is administered intranasally in accordance with the above aspectsbut in a dose or doses adding up to between about 15 nanograms and about10 milligrams.

Whether administered intranasally or otherwise, rhCC10 can be givenalone, in conjunction with, before or after other standard rhinitis andsinusitis treatments, including but not limited to intranasal orsystemic corticosteroids, NSAIDs (including aspirin, COX-2 inhibitors),pain medications, antibiotics, antivirals, antifungals, decongestants,antihistamines, chromium solutions, nasal lavage, saline nasal lavage,and homeopathic remedies.

In another aspect, rhCC10 can be used as an excipient and/or localanti-inflammatory, and/or local immunosuppressor, to facilitate thelocal nasal delivery or application for local delivery or systemicabsorption of other drugs to the nasal tissues that may or may notirritate or otherwise elicit, or may elicit, an undesired localirritation at the site of application. Thus, rhCC10 may be used as anexcipient for other drugs to alleviate or avoid discomfort associatedwith nasal delivery.

In another aspect, rhCC10 can be used as an excipient or to alleviatethe irritation caused by intranasal administration of other drugs foreither local or systemic delivery.

In addition, rhCC10 can be formulated as an aqueous solution, asuspension (containing a nasal surfactant excipient), or a gel (such asa hydrogel employing, for example, hydroxymethylcellulose), in order toachieve the proper viscosity for nasal application and localdistribution profile in the nasopharyngeal cavity. Likewise, rhCC10 canbe formulated in combination with other active ingredients such asantibiotics or other antimicrobial agents, saline nasal lavages,decongestants, mucolytics, LTRA's, β-agonists, bronchodilators, etc.

In another aspect, rhCC10 is formulated as an aqueous solution that isloaded into a nasal spray squeeze bottle, metered dose inhaler or spraypump device. In yet another aspect, rhCC10 is formulated as a suspensionin a surfactant that is loaded into a nasal syringe-type applicationdevice, a metered dose inhaler, or other nasal application device.Further, in still another aspect, rhCC10 is formulated in a hydrogel, orother form of artificial mucus, and single doses are placed in a singleuse nasal swab device for intranasal application.

DETAILED DESCRIPTION

The present invention relates to the critical dosages and timing ofadministration of rhCC10 to treat, cure or prevent nasal rhinitis andsinusitis, especially chronic nasal rhinitis with recurrent sinusitis,chronic rhinosinusitis, and nasal polyposis in humans. The rhCC10 ispreferably obtained by the processes described in U.S. PatentApplication Publication Nos. US 2003-0109429 and US 2003-0207795attached hereto at Ex. A & B, respectively, both of which areincorporated by reference in their entirety, or via any other processwhich yields pharmaceutical grade (meeting FDA requirements) rhCC10. TherhCC10 of the embodiments of the present invention can be administeredwith, without, before or after other intranasal, pulmonary, or systemictherapy.

Dosages

Preferably, in treating or preventing nasal rhinitis, sinusitis, chronicrhinosinusitis, and nasal polyposis, rhCC10 is administeredintranasally, to each nostril 1-3 times per day, for 7-14 days, andevery other day thereafter for another 14 days, and thereafter asneeded. More preferably, rhCC10 is administered as soon as the patientbegins to experience sinus pain and pressure.

To effectuate the desired outcomes which are further described below,reference is made to methods of administration described in thefollowing embodiments:

In one embodiment, a dose or multiple doses of rhCC10 equaling a doseranging from about 1.5 micrograms to about 1.5 milligrams can beadministered. In another embodiment, rhCC10 can be administered in thedose range on a daily basis. In yet another embodiment, rhCC10 can beadministered in the dose range on a daily basis for at least seven daysconsecutively. In still a further embodiment, rhCC10 can be administeredin the dose range on a daily basis for at least 14 days consecutively.In still another embodiment, rhCC10 can be administered in the doserange every other day for 30 days consecutively. In yet anotherembodiment, rhCC10 can be administered in tapered dosages daily for tenconsecutive days, said tapered dosages comprising a high dose at eachadministration for the first three days, an intermediate dose at eachadministration for the second three days, and a low dose at eachadministration for the last four days. In yet still another embodiment,rhCC10 can be administered in the dose range or in tapered doses up tothree times per day, approximately every eight hours.

In another embodiment the above doses of rhCC10 can be administeredintranasally to the patient. In yet another embodiment, the above dosesof rhCC10 can be administered to the patient as an aerosol, byintranasal instillation, or by deposition of a gel or cream in nasalpassages. In a further embodiment, rhCC10, in accordance with themethods described above, can be administered prior to, during or afteran oral or intranasal decongestant, anti-histamine, corticosteroid,mucolytic, expectorant, mucus suppressor, surfactant, bronchodilator,vasoconstrictor, sinus pain analgesic, or other typical therapy. Instill another embodiment, rhCC10, in accordance with the methodsdescribed above, can be administered to treat or prevent nasal rhinitis,nasal sinusitis, chronic rhinosinusitis, or nasal polyposis in apatient.

The doses of rhCC10 and application methods described above can beadministered daily, more than once daily, three times daily, every otherday or in a tapered fashion depending upon the severity of disease beingtreated, the patient's overall health, and whether an acute or chroniccondition is being treated. For example, the more severe the diseasecondition, the higher the amount of rhCC10 would be required toeffectively treat the disease. For maintenance therapy of chronicdisease, for example, to prevent an exacerbation of nasal rhinitis,nasal sinusitis, or nasal polyposis, lower doses would be used. It isunderstood that a physician would be able to monitor and adjust doses,formulations, and application methods as needed based on the patient'ssymptoms and responses to therapy and within the parameters and doseranges described in the embodiments of the present invention.

Formulations

rhCC10 is maximally effective when applied directly to the local nasalepithelium, for example by use of a liquid formulation in a spraybottle, spray pump, or lavage. Therefore, it is sometimes necessary touse fast-acting local mucolytics, anti-histamines, and/or decongestants,as well as physical methods such as inhaling moist warm air, hotcompresses applied to the face, and salt water nasal lavage to open upthe nasal passages before rhCC10 can be applied effectively to the nasalepithelia.

Intranasal instillation is another method for administering rhCC10 thatcan be accomplished using rhCC10 in either a liquid or gel formulation.The gel dosage formulation provides the advantage of better local dosingover a longer period of time by retaining the dose of rhCC10 in thelocal nasal area in which it was swabbed longer, whilst liquid dosagesmay be partially swallowed following instillation due to normal nasaldrainage resulting in much shorter local exposures and smaller localdoses. However, intranasal instillation of a liquid dosage form by nasallavage, using a “neti pot” type of device, confers the advantage thatthe dose is more immediately distributed over a larger surface area inthe nasal tissues and sinuses, than a local application gel formulation.

rhCC10 can be formulated with several nasal excipients for intranasaldelivery. These include excipients to adjust the pH of the drug, tobuffer the drug to maintain solubility, to act as preservatives orenhance preservatives for prevention of microbial growth and/ortransfer, to adjust the tonicity, solubility, or viscosity of the drug,to enhance penetration or permeation of the drug (systemic delivery), tomodify the local bioavailability and half-life of the drug (increaseviscosity), to reduce toxicity, to suspend insoluble drugs, and to alterthe taste of the formulation. Table 2 contains a non-exclusive list ofexemplary excipients and their functions in intranasal formulations ofrhCC10. Any single excipient or combination of excipients can be used toformulate rhCC10 for intranasal administration.

TABLE 2 Examples of Excipients for Intranasal Formulations ExcipientFunction Acids (hydrochloric, acetic, citric) pH adjustment, bufferSodium hydroxide pH adjustment Sodium & potassium salts (Sodium Bufferacetate, sodium citrate, sodium phosphate, potassium phosphate) Edetatedisodium Preservative enhancer, Metal chelator Benzalkonium chloridePreservative Benzethonium chloride Preservative Benzyl alchohol (akaphenylcarbinol, Preservative etc.) Chlorobutanol PreservativeMethylparaben Preservative Phenylethyl alcohol PreservativePhenylmercuric acetate Preservative Propylparaben PreservativeThimerosal Preservative Sodium or potassium chloride Adjust tonicity(make isotonic) Microchrystalline cellulose Adjust viscosity Nacarboxymethylcellulose Adjust viscosity Hydroxyethylcellulose Adjustviscosity Ethanol Solvent Glycerol Solvent/Adjust tonicity GlycineSolvent/Adjust tonicity Dextrose Adjust tonicity Polyethylene glycol(PEG) Solvent Propylene glycol Solvent Glyceryl dioleate SolventGlyceryl monoleate Surfactant/emulsifier (suspend lipophilic drugs)Lecithin Surfactant/emulsifier (suspend lipophilic drugs) Polysorbate 20& 80 (aka Tween Surfactant/emulsifier (suspend 20 & 80) lipophilicdrugs) Triglycerides Multiple Menthol Modify flavor Saccharin sodiumModify flavor Sorbitol Modify flavor Chitosan Permeation enhancerCyclodextrin Permeation enhancer Bile salts Permeation enhancerLiposomes Permeation enhancer Starch microspheres Permeation enhancerGlycerrhizin Permeation enhancer

rhCC10 can also be formulated in combination with other drugs,artificial mucus, or other active ingredient for intranasaladministration. Drugs with which rhCC10 can be formulated for intranasaladministration include, but are not limited to, local or systemicantimicrobial agents (antivirals, antibacterials, antifungals),decongestants, anti-histamines, mucolytics, expectorants, leukotrienereceptor antagonists, bronchodilators, beta₂-adrenergic receptoragonists, local-acting vasoconstrictors (such as oxymetazoline),anti-inflammatory agents, and analgesic agents. Still other drugs withwhich rhCC10 can be formulated for intranasal administration for localor systemic effects include anti-inflammatory agents, beta₂-adrenergicreceptor agonists, anti-cancer agents, anti-angiogenic agents,anti-fibrotic agents, immunomodulatory agents, vaccines, metabolicagents, analgesics, neuroleptic agents, anesthetics, agents fordepression and other psychiatric disease (mental health), anti-addictionagents, homeopathic remedies, herbal preparations, vitamins and mineralsand the like.

rhCC10 is compatible with most non-reactive chemicals and drugs,including hydrophilic and hydrophobic chemicals, nucleic acids andnucleic acid analogs, proteins and peptides, carbohydrates, lipids andphospholipids, etc. As a secretoglobin critically involved in transportof substances in epithelial cells, rhCC10 is ideally suited to enhancethe delivery of other drugs via the nasal passages. rhCC10 can also actas a local anti-inflammatory agent that can be used as an excipient tosuppress painful local nasal responses at the site of administration ofother drugs, such as, for example, chemotherapeutic agents and drugsthat produce a “burning sensation” upon administration.

A key parameter relevant to drug efficacy associated with intranasallyadministered rhCC10 is the concentration of the rhCC10 itself.Formulations in which the rhCC10 concentration is too high (i.e. above 2mg/ml) have demonstrated null or even detrimental effects, as evidencedby the clinical outcomes described in the Background. The rhCC10formulation was 5.6 mg/ml and was applied directly to the patient'snostrils (Widegren, et al. 2009). Conversely, in example 4, in which therhCC10 concentration was 250-262 micrograms/ml, a clinical benefit wasconferred. In an unrelated experiment, pre-term lambs treated with 5mg/kg of body weight using a 5.5 mg/ml formulation of rhCC10,administered by intratracheal instillation, suffered from severe hypoxiaand ¾ animals died of respiratory failure within four hours of drugadministration, while none of the four placebo treated animals died(unpublished; Ikegami, M., Univ of Cincinnati). In contrast, when thesame formulation of rhCC10 was diluted to 2 mg/ml and administered viaintratracheal instillation to intubated pre-term lambs, the animalsshowed various benefits from receiving the drug compared to placebo(Miller, 2005a, 2005b, 2007; Shashikant, 2005). Without being bound toany particular theory, the phenomenon may be related to the very highsphere of hydration for rhCC10, that is, the number of water moleculescoordinated by CC10 is higher than the average protein. Administrationof high concentrations of CC10 to mucosal and other bodily fluids mayresult in the “subtraction” or loss of water from the local fluids,thereby causing a local dehydration and detrimental disruption in theequilibria between substances in the local biological milieu.Alternatively, or in conjunction with the preceding, an acute localover-abundance of CC10 may also result in desensitization of cells andtissues to the presence of CC10, effectively reducing the potency of thedrug rather than increasing the pharmacologic effect. In some cases,feedback inhibition of a pathway or set of pathways involving aparticular metabolite or mediator may actually result in the oppositeeffect (activation versus suppression and vice versa) than may beobserved at lower doses of the metabolite or mediator. A cutoff forrhCC10 formulations intended to be administered to nasal and othermucosal surfaces by intranasal, intratracheal or other local/topicaladministration is 2 mg/ml, above which rhCC10 is not efficacious and caneven be detrimental.

The following detailed examples are illustrations of embodiments. Itshould be clear that these are not intended to limit the scope of thepresent invention.

Example 1 Intranasal Administration of rhCC10 to Allergic RhinitisPatients

RhCC10 was produced in E. coli bacteria and purified by a process(Claragen, Inc., College Park, Md.), described in U.S. ApplicationPublication Nos. US 2003-0109429 and US 2003-0207795, both of which areincorporated by reference in their entirety. The protein for the studywas provided as a >98% pure solution of recombinant human CC10homodimer. The biological activity of each batch was compared using aproprietary secretory PLA₂ inhibition assay, described in U.S.Application Publication Nos. US 2002-0169108 which is incorporatedherein by reference.

In the nasal allergen challenge model, patients with known seasonalallergies to known allergens were subjected to instillation of anallergen solution into the nasopharyngeal cavity on seven consecutivedays. In order to minimize safety risks the amount of allergen instilledwas carefully calibrated in each patient to elicit a mild local allergicresponse that is quantitated using four main outcome parameters over theseven day challenge period. These parameters include; 1) total nasalsymptom scores, 2) peak nasal inspiratory flow, 3) quantitation ofbiomarkers in nasal lavages, and 4) response to histamine challenge.

A total of 35 patients with seasonal allergies to tree pollens completeda placebo-controlled, randomized, blinded, cross-over nasal allergenchallenge study of rhCC10 versus placebo at the Lund University Hospitalin Lund, Sweden. The purpose of the study was to determine the safety,tolerability and efficacy of intranasal administration of recombinanthuman Clara Cell 10 kDa (rhCC10) protein in subjects with allergicrhinitis. In order to investigate whether intranasal administration ofrhCC10 could mitigate the nasal rhinitis symptoms caused by nasalallergen challenge.

Patient responses to nasal allergen challenges in the absence of rhCC10were first measured and baseline data recorded. A total of 39 patientswere screened for inclusion in the study. All patients were malesubjects, aged 18-50 years, with Body Mass Index between 18 and 28kg/m², and a history of birch and/or timothy pollen-induced seasonalallergic rhinitis for at least the previous 2 years and otherwisehealthy. Each patient had elevated specific IgE or at least one positiveskin prick test (SPT) to at least one aero allergen (eg. timothy orbirch pollen) and each patient exhibited symptoms provoked by theallergen with a corresponding elevated specific IgE or positive SPT.Subjects were excluded from the study if they had perennial allergy(e.g. chronic rhinitis), except for cat and/or dog sensitivity under thecondition that these subjects are not exposed to cats and dogs. Subjectswere also excluded if they had other nasal disease (eg. structuralabnormalities of the nose, rhinosinositis, or nasal polyposis), anyupper respiratory tract infection during the period of 2 weeks beforethe start of the study, were currently receiving treatment or hadreceived treatment within 4 weeks of enrolment with intranasal, inhaledor systemic glucocorticosteroids, β2-adrenergic receptor agonists, orany other anti-inflammatory medication, or had a bacterial or fungalinfection within the past month prior to enrollment. A summary ofpatient characteristics and baseline data is given in Table 2.

TABLE 2 Summary of Demographic and Baseline Data Age (years) Mean (SD)26.1 (5.5) Range 19-48 Race Caucasian 39 Gender Male 39 Body weight (kg)Mean (SD) 81.1 (10.3) Height (cm) Mean (SD) 182.6 (7.2) Body mass index(BMI) Mean (SD) 24.3 (2.53) Systolic blood pressure Mean (SD) 121.4(9.2) Diastolic blood pressure Mean (SD) 77.4 (5.9) Pulse rate Mean (SD)68.8 (8.2) Ear, nose and throat Normal 38 Abnormal 1 CardiovascularNormal 39 Respiratory Normal 39

Patients were initially screened for their responses to tree pollenallergens to determine the amount of allergen administered during thechallenge periods which was calibrated based on individual patientresponses. A physical examination was given, including ear, nose andthroat examination and vital signs. Then a nasal allergen challenge wasperformed in order to establish the allergen dose which resulted in atleast 5 sneezes and/or a symptom score of at least 2 on a scale of 0-3for either the symptoms of nasal congestion or rhinorrea. Rhinorrea isdefined as a discharge from the nasal mucus membranes and is typicallywatery. All allergen administrations were performed in the clinic byhospital staff. In order to estimate, for each individual, thesymptom-producing, tolerable, and repeatable allergen dose for the nasalchallenge series, a titration procedure was performed in all subjects.The allergen (birch or timothy pollen) that induced the most significantwheal reaction in the skin prick test was chosen for the nasaltitration. Increasing doses of allergens (Alutard™, ALK, Denmark) wasadministered at 10-min intervals using a nasal spray-device. Thespray-device delivered 100 μl per actuation, and one puff was sprayedinto each nostril resulting in effective doses of 100, 300, 1000, and3000 SQ-Units per nasal cavity. This scheme was followed until thepatient responded acutely with at least five sneezes and/or a symptomscore of at least 2 or more on a scale from 0 to 3 for either of thesymptoms nasal blockage and runny nose. The allergen dose that producedthis effect was chosen for the daily allergen challenge series duringthe first and second treatment periods (eg. cohorts.) The type andamount of allergen administered to each patient is shown in Table 3.

TABLE 3 Selection of allergen and number of sneezes at allergentitration per subject Selection of allergen (Histamine/ 100 300 10003000 Subject Birch ) SQ-Units SQ-Units SQ-Units SQ-Units 1 Timothy 6 — —— 2 Timothy 3 — — — 3 Timothy 0 0 0 0 4 Timothy 1 1 4 6 5 Timothy 0 2 —— 6 Timothy 0 0 0 — 7 Timothy 1 5 — — 8 Timothy 3 4 0 — 9 Birch 0 4 — —10 Timothy 10 — — — 11 Timothy 0 — — — 12 Timothy 0 3 — — 13 Timothy 0 0— — 14 Birch 3 7 — — 15 Timothy 0 1 — — 16 Timothy 1 — — — 17 Timothy 25 — — 18 Timothy 3 0 — — 19 Birch 0 0 0 — 20 Timothy 0 0 4 — 21 Timothy4 4 — — 22 Timothy 3 — — — 23 Birch 1 10 — — 24 Birch 3 — — — 25 Birch 44 — — 26 Birch 0 — — — 27 Timothy 4 2 — — 28 Timothy 0 0 0 — 29 Birch 00 0 1 30 Timothy 9 — — — 31 Timothy 11 — — — 32 Timothy 1 1 2 — 33 Birch0 0 2 — 34 Timothy 2 2 — — 35 Timothy 6 — — — 36 Timothy 1 7 — — 37Timothy 0 3 — — 38 Birch 0 5 — — 39 Timothy 0 0 3 —

The study was performed during the pollen-free winter months. The studywas blinded and placebo-controlled in that doctors and patients were notaware of whether they were receiving rhCC10 or placebo. The study wasrandomized in that patients were randomly assigned to either the rhCC10or placebo treatment groups. The study was cross-over design in thateach patient was treated in two seven day cohorts separated by a washoutperiod of 3-5 weeks. Each patient completed a cohort in which theyreceived rhCC10 and one in which they received placebo. Patients wereallowed to take several types of non-steroidal medications, as needed,to relieve their nasal and sinus discomfort during the treatment period.Analgesics (including aspirin but not ibuprofen) and antibiotics wereallowed, and Clarityn® (Claritin™) 10 mg, was allowed in case of severeallergic symptoms and was provided by the clinic.

The test agents, placebo and rhCC10, were placed in 10 ml glass vials,labeled numerically so that doctors and patients in the clinic would notbe able to distinguish between them. A key was maintained in thehospital pharmacy and doctors were to be informed of the identity ofeach vial only in the event of an adverse event in which the doctorwould need the information to treat the patient. A disposable medicalnasal spray device, manufactured by Valois Pharm (France), was connectedto the 10 mL vials at the clinical site just before administration. Thisdevice consisted of a pump (VP7/100S 18PH), an actuator (PR147) and acap (B25/A). Placebo consisted of sterile, unbuffered 0.9% sodiumchloride. The rhCC10 was in sterile unbuffered 0.9% sodium chloride at aconcentration of 5.6 mg/ml. Both placebo and rhCC10 appeared as clear,colorless, odorless liquids that could not be readily distinguished. Atotal of 100 microliters of placebo or rhCC10 was administered to eachnostril of each patient on each day of treatment for a total of sevenconsecutive days of treatment in each treatment period. All allergen andtest agent administrations were performed in the clinic by hospitalstaff. The total daily dose of rhCC10 was 1.1 milligrams per day,administered in a single dose as an aerosol sprayed in a 100 microlitervolume to each nostril, or 0.56 milligrams per nostril. The rhCC10 wasadministered 15′-30′ prior to administration of allergen.

The outcomes were measured as follows:

1. Total Nasal Symptom Score (TNSS)

Nasal symptoms, including nasal congestion, rhinorrea and sneezy/itchynose were scored by the patients and recorded in the patient diary priorto administration of study medication in the morning (rating symptomsduring the preceding 12 h, but disregarding possible symptoms the first15 minutes post study medication). TNSS was recorded 15 minutes aftereach allergen challenge. In addition, symptoms were scored in theevening (again reflecting symptoms, during the preceding 12 h, exceptingsymptoms directly post dosing). The symptoms were each scored accordingto the following: 0=no symptoms, 1=mild symptoms, 2=moderate symptoms,3=severe symptoms. The scores were added to constitute a total score,per time point, which ranged from 0 to 9. Mean nasal symptom scores, formorning recordings, for recordings 10 minutes after allergen challenge,and for evening recordings, respectively, of the last three days of eachallergen challenge period was used in the statistical analysis.

2. Peak Nasal Inspiratory Flow (PNIF)

PNIF was measured by the patients before the intake of the drug in themorning, 10 minutes after the allergen challenge, and in the evening.The measurements were carried out using a PIF-meter (Clements-Clarke,Harlow, U. K.) equipped with a facial mask. Patients stood up during theprocedure, placed the mask snugly over the face with both hands, closedthe mouth and inhaled through the nose. They recorded the value andreturned the device to a reading of 30, then repeat the procedure 2 moretimes. The highest value of the three measurements was recorded in thediary. Similar to the nasal symptom score, PNIF recordings, per timepoint, of the last three days of each allergen challenge period wereused in the statistical analysis.

Example 2 Co-Administration of Medications

A total of ten patients in both the placebo and rhCC10 groups requiredrescue medications to treat their discomfort and allergy symptoms whileon the protocol. As can be seen from Table 5, a variety of local andsystemic drugs for relief of intranasal symptoms were taken, includinganti-inflammatory agents, allergy medications, anti-histamines,corticosteroids (Flutide), and oxymetazoline.

TABLE 5 Co-administration of rescue medications in patients receivingplacebo or rhCC10 Subject Treatment Day Medication (trademark/INN) 3rhCC10 2 Clarityn ® (Claritin ™)/loratadin 6 Clarityn ®(Claritin ™)/loratadin 7 Neseril/oximetazolin 3 placebo 4 Clarityn ®(Claritin ™)/loratadin 5 rhCC10 5 Loratadin/loratadin 6 placebo 2Alvedon/paracetamol 12 placebo 3 Loratadin/loratadin 13 placebo 3Loratadin/loratadin 15 placebo 3 Loratadin/loratadin 18 placebo 5Clarityn ® (Claritin ™)/loratadin 6 Loratadin/loratadin 25 rhCC0 1Alvedon/paracetamol 5 Loratadin x2/loratadin 7 Flutide Nasal/flutikason25 placebo 7 Antihistamin intake after evening PNIF 26 rhCC10 5Loratadin/loratadin 6 Loratadin/loratadin 38 rhCC10 5 Clarityn ®(Claritin ™)/loratadin 7 Kestine/ebastin

Loratadin is a non-sedating antihistamine, paracetamol, is an analgesicand antipyretic, flutikason (fluticasone) is a corticosteroidanti-inflammatory agent, oxymetazoline is a selective alpha-1 agonistand partial alpha-2 agonist topical decongestant, and ebastin is anon-sedating H₁ antihistamine. These patients did not experience anysignificant AEs as a result of co-administration of these other agentssimultaneously with the rhCC10, therefore, the combination of rhCC10with these drugs is safe and pharmacologically-acceptable.

Example 3 Safety and Tolerability of Intranasal Administration of rhCC10

As part of the safety assessment for this proof of concept intranasaladministration of rhCC10 in humans adverse events (AEs) and seriousadverse events (SAEs) were monitored, recorded and reported. Theclinical investigator was responsible for the detection anddocumentation of events meeting the criteria and definition of an AE orSAE. An AE is any untoward medical occurrence in a subject or a clinicalinvestigation temporally associated with the use of the investigationaldrug whether or not the event is considered to have a causalrelationship with the drug. In this trial, a pre-existing condition(i.e., a disorder present before the AE reporting period started andnoted on the pre-treatment medical history/physical examination form)was not reported as an AE unless the condition worsened or episodesincreased in frequency during the AE reporting period. Serious adverseevents were defined as any untoward medical occurrence that, at anydose; 1) results in death, 2) is life-threatening, 3) requireshospitalization or prolongation of an existing hospitalization, 4)results in disability/incapacity, 5) is a congenital anomaly/birthdefect, 6) is an important Other Medical Event (OME), and 7) all grade 4laboratory abnormalities. The AE reporting period for began uponreceiving the first dose of investigational medication and ended at the2-week post discontinuation of investigational medication visit(follow-up visit).

No SAE's occurred during the study. Overall, a total of 15 adverseevents were reported in subjects in both the placebo and rhCC10treatment groups. All AEs were rated as mild in severity. In each group,11 of 15 AEs were rated as non-assessable with respect to relatedness tostudy drug while four of 15 AEs in each group were rates as unlikely tobe related to study drug. A summary of AEs for each patient receivingplacebo is given in Table 6 and for those receiving rhCC10 at the timeof the AE are given in Table 7.

TABLE 6 List of adverse events for patient receiving placebo ReportedPatient Maximum as Relationship to number Description intensity serious?trial drug 6 Headache 1 = mild 0 = No 1 = unlikely 12 Gastric influenza1 = mild 0 = No 4 = not assessable 12 Gastric influenza 1 = mild 0 = No4 = not assessable 15 Ear pain 1 = mild 0 = No 1 = unlikely 15 headache1 = mild 0 = No 4 = not assessable 15 fatigue 1 = mild 0 = No 4 = notassessable 15 ear pain 1 = mild 0 = No 4 = not assessable 20 Sorethroath 1 = mild 0 = No 4 = not assessable 20 Common cold 1 = mild 0 =No 4 = not assessable 25 Headache 1 = mild 0 = No 1 = unlikely 26 Sorethroat 1 = mild 0 = No 4 = not assessable 27 stomach ache 1 = mild 0 =No 1 = unlikely 29 common cold 1 = mild 0 = No 4 = not assessable 31Fever 1 = mild 0 = No 4 = not assessable 38 urticaria 1 = mild 0 = No 4= not assessable

TABLE 7 List of adverse events for patient receiving rhCC10 Report-Patient Maximum ed as Relationship to number Description intensityserious? trial drug 1 Common cold 1 = mild 0 = No 4 = not assessable 2Common cold 1 = mild 0 = No 1 = unlikely 2 Common cold 1 = mild 0 = No 1= unlikely 7 Sore throat 1 = mild 0 = No 1 = unlikely 16 fatigue 1 =mild 0 = No 4 = not assessable 16 fatigue 1 = mild 0 = No 4 = notassessable 23 Headache 1 = mild 0 = No 4 = not assessable 23 Common cold1 = mild 0 = No 4 = not assessable 26 Common cold 1 = mild 0 = No 4 =not assessable 28 tired 1 = mild 0 = No 4 = not assessable 28 tired 1 =mild 0 = No 4 = not assessable 28 headache 1 = mild 0 = No 4 = notassessable 32 Headache 1 = mild 0 = No 4 = not assessable 38 ague 1 =mild 0 = No 4 = not assessable 39 Mild cold 1 = mild 0 = No 1 = unlikely

Therefore, intranasal rhCC10 administration was found to be safe andwell-tolerated in humans when given once daily as an aerosol in adivided dose of 1.1 milligrams, 0.56 milligrams per nostril, for sevenconsecutive days.

Example 4 Intranasal Administration of rhCC10 to a Patient with ChronicRhinitis and Recurrent Sinusitis (Aka Chronic Rhinosinusitis)

An 11 mg (2 mls) aliquot of rhCC10 was added to a soft plastic squirtbottle containing 42 mls of sterile 0.65% saline containing disodium andmonosodium phosphate, and phenylcarbinol (preservative) plusbenzylkonium chloride (preservative), or 0.1% thimerosol (also apreservative), creating a 250 microgram/ml solution of rhCC10. Thepatient self-administered the rhCC10 by inserting the applicator end ofthe bottle to the nose, such that the aperture that dispenses the drugis held inside the nostril, and simultaneously squeezing and inhaling. Asimple aerosol is created when the bottle is rapidly squeezed, forcingliquid through a small pinhole at the top of the nasal applicator end.The volume and dose delivered depends upon the rapidity of the squeezeand force exerted. Volumes ranging from 25-500 microliters,corresponding to 6.6-131 micrograms of rhCC10, are typically dispensed.When the squeeze is harder and larger volumes are delivered, the nasalpassages are lavaged and part of the dose may be swallowed or flow intothe trachea over a period of several minutes.

In this example, rhCC10 was administered to a patient suffering fromepisodic and/or chronic sinus pain due to chronic rhinosinusitis,stemming from perennial allergies, with recurrent bacterial sinusinfections. The patient's history includes; 1) antibiotics prescribedfrom two to twelve times per year for sinus infection in the past sixyears, 2) intranasal corticosteroids prescribed and taken as needed forthe past six years, and 3) non-prescription analgesics, decongestantsand anti-histamines taken daily to relieve sinus pain, nasal and chestcongestion, and enable the patient to sleep through the night. RhCC10was a useful substitute and adjunctive therapy for the patient in thefollowing doses, dosing regimen, formulations, and drug-devicecombinations.

In the first dosing regimen, the patient suffered from a painfulbacterial sinus infection for three days prior to self-administering therhCC10 from the squirt bottle, with two squirts per nostril, three timesper day for three days. The total daily dose intake using this methodranged from 78.6-1,572 micrograms total (39.3-786 micrograms pernostril), corresponding to 1.1 micrograms/kg-22.5 micrograms/kg of bodyweight per day in the average 70 kg patient. The patient then taperedthe dosing down to twice per day for two days (52.4-1,024 microgramsdaily total (26.2-512 micrograms per nostril), corresponding to 749nanograms/kg-14.6 micrograms/kg per day in the average 70 kg patient),and then to once per day for two days (26.2-524 micrograms daily total,corresponding to 374 nanograms/kg-7.5 micrograms/kg per day in theaverage 70 kg patient). After one week of tapered dosing regimen withintranasal rhCC10, the patient discontinued use. The patient's sinuspain, rhinitis and bronchitis symptoms (nasal congestion, sneeze, cough,airway constriction and chest congestion), and sleeplessness,disappeared within 24 hours of initiation of treatment and did notreturn for at least six weeks following the final dose of rhCC10.Intranasal rhCC10 was safe and well tolerated in the patient, althoughsome drying of nasal mucous membranes was experienced.

Example 5 Intranasal Administration of rhCC10 to a Patient to PreventRecurrent Sinusitis

In a second dosing regimen, the patient received rhCC10 in theformulation and spray bottle of Example 4, twice per day, in the morningand evening, starting within 12 hours of sensing the first sinus pain.The sinus pain was associated with the recurrence of a bacterial sinusinfection that had been treated for 14 days with a powerfulbroad-spectrum antibiotic (eg. Levaquin), during which time the painabated, but returned within four days of ending the antibiotic. Knownside effects associated with the antibiotic occurred in the patient,including constipation and irritable bowels, chest pain, dizziness,transient numbness and tingling in the extremities, extreme sunburn, andincreased susceptibility to bruising. As a result, the doctor advisedthe patient to avoid further use of the antibiotic. The patient thenused rhCC10 with over-the-counter decongestants to treat the symptomsassociated with the recurrent sinus infection. Within 24 hours of thefirst dose, the nasal pain and congestion disappeared. The patientcontinued to administer the rhCC10 twice per day for one week, thendecreased to once per day for one week, then discontinued therapy. Thebacterial infection did not recur for at least six weeks followingintranasal rhCC10 therapy.

Example 6 Intranasal Administration of rhCC10 for Maintenance

Maintenance therapy with rhCC10 to prevent sinus pain and infection,often arising from seasonal or perennial allergy and exposure toairborne allergens, is also possible. Daily intranasal administration ofrhCC10, at a formulation concentration not to exceed 500 micrograms/ml(preferably not to exceed 250 micrograms/ml), in doses of 26.2-524micrograms total, given in single or multiple actuations per nostril,corresponding to 374 nanograms/kg-7.5 micrograms/kg of body weight) forup to two and one half months would safely control chronic rhinitissymptoms, rhinosinusitis, nasal and chest congestion, sinus infectionand pain, and sleeplessness, and prevent the need for antibiotics,analgesics (NSAIDS such as aspirin, ibuprofen), decongestants,anti-histamines, and sleep-inducing drugs.

Using these methods, formulation, dose, dosing regimen, and drug-devicecombinations, rhCC10 was efficacious in the alleviation of symptomsassociated with chronic rhinitis and bacterial sinus infection (akachronic rhinosinusitis). In still other instances of severe or recurrentsinus infection, several other antibiotics (Amoxicillin, Zithromax,Biaxin, etc.) were used to contain bacterial growth while rhCC10alleviated the pain and symptoms. For mild infections and to preventsevere painful infections, rhCC10 was used without an antibiotic, thus,sparing the patient the negative side effects associated with theantibiotic. No adverse events were associated with potentialinteractions between rhCC10, decongestants, antihistamines, andantibiotics. Thus, over the counter decongestants and antihistamines andantibiotics commonly prescribed for nasal sinusitis were either avoidedentirely or used safely in conjunction with rhCC10 to alleviate moderateto severe nasal symptoms.

Example 7 Intranasal Administration of rhCC10 for Treatment ofCorticosteroid-Refractory, Antibiotic Resistant, Acute Sinus Infection

The patient suffered from a severe ongoing sinus infection,characterized by pain, pressure, disruption of sleep, loss of bloodpressure upon standing, and inability to walk. The patient had noallergies to airborne allergens (seasonal or perennial) and theinfection occurred in the month of January when no seasonal allergenswere present. The diagnosis and severity of the sinus infection wasverified by CT scan. Prior to receiving rhCC10, the patient had been onantibiotics for 5 weeks (amoxicillin; 500 mg/day; 10 days; thenaugmentin, 4 grams/day for 3 weeks) and on intranasal corticosteroidtreatment for 10 days (fluticasone propionate). Despite thesetreatments, he remained in considerable pain, with pressure throughouthis sinuses, and facial edema (puffiness). Immediately prior toreceiving rhCC10, both sides of his nasal septum were blood red andcontained readily visible dilated capillaries, indicating the presenceof severe local inflammation. A 250 microgram/ml solution of rhCC10 in0.65% saline containing disodium and monosodium phosphate,phenylcarbinol (preservative) and benzylkonium chloride (preservative)was then administered in a single intranasal dose as a spray into eachnostril at a dose of approximately 20-50 micrograms per nostril.Approximately 12 hours after receiving the single intranasal dose ofrhCC10, his nasal septum was a normal dusky purple with no dilatedcapillaries visible, indicating a profound local anti-inflammatoryeffect. The patient continued on rhCC10, twice daily, for 7 days, notingdecreased sinus pain and pressure symptoms. The dosing regimen of rhCC10was to be tapered from two squirts per nostril, twice per day for 3days, to one squirt per nostril twice per day for 3 days, to one squirtper nostril once per day for 3 days. The patient continued with thisregimen for 4 days. However, it was noted on day 5 that the patientstill had intense pain in the ethmoid sinus, which is difficult toaccess with a nasal spray. Thereafter, the rhCC10 was administered onthe same schedule by a lavage technique to increase access of rhCC10 tothe surfaces of the ethmoid sinus region. In the lavage method, a totaldose of 250 micrograms of rhCC10 (i.e. 1 ml of the 250 microgram/mlsolution) was added to 118 mls (½ cup; 4 fluid ounces) of a standardcommercially available nasal lavage solution. The patient received thelavage in the supine position with head tilted back, allowing the rhCC10formulation to settle in the sinuses for 3-5 minutes. The patient thensat up, allowing the lavage to flow out and be expelled by nose blowing.The lavage was administered twice per day for 2 days, then once per dayfor 3 days, then discontinued. A CT scan performed 21 days after theinitial dose of rhCC10 revealed complete resolution of the sinusinfection without evidence of scarring, epithelial thickening, or otherremaining blockage. The rhCC10 formulation mediated a potentanti-inflammatory response, which was caused by a bacterial infectionand not allergy. rhCC10 further mediated an anti-inflammatory responsewhen standard anti-inflammatory therapy in the form of intranasalcorticosteroids failed. rhCC10 also facilitated clearance of thebacterial infection, which resolved without the use of additionalantibiotics. Finally, rhCC10 mediated a complete recovery of the nasalepithelia, avoiding the scarring, fibrosis, and epithelial thickeningthat typically accompanies such severe infections.

Example 8 Intranasal Formulation of rhCC10

Intranasal delivery of rhCC10 is useful for example, when treating upperrespiratory (nasal and sinus and upper airway) inflammation andfibrosis, due to perennial allergy, infection, or some other form ofacute or chronic upper respiratory irritation. rhCC10 is soluble in awide range of aqueous solutions, over a wide range of pH values, forexample 3.9-8.5, and in a wide range of salt concentrations (for example0.1%-4%), as well as a variety of alcohol/water mixtures (for example0.1%-90% ethanol). Thus, rhCC10 has the solubility and stabilitycharacteristics, to be used with a wide range of intranasal dispensingdevices, including, but not limited to, for example, simple squirtbottles with uncontrolled volumetric doses for self-administration ofliquid aerosols, pump-action or pressurized canister metered dosedevices for self-administration of liquid aerosols, propellant-drivendry powder or liquid aerosol metered dose devices forself-administration, gel-laden nasal swabs for topical delivery to thenasal passages, and drug-loaded syringes for deeper topicaladministration and sinus lavage, for voluntary or involuntaryadministration to the conscious or unconscious patient.

Based on the foregoing, the critical ranges for rhCC10 dosages effectiveto safely treat, cure and prevent nasal rhinitis, especiallynon-allergic rhinitis, nasal sinusitis, chronic rhinosinusitis, andnasal polyposis have been found. Accordingly, the present inventionprovides a safe and well-tolerated intranasal rhCC10 based therapyeffective at treating the symptoms of nasal rhinitis, especiallynon-allergic rhinitis, nasal sinusitis, chronic rhinosinusitis, andnasal polyposis thus reducing the significant morbidities in child andadult patients suffering from these conditions, while not causing anydangerous side effects.

1. A safe and well-tolerated method of treating rhinosinusitis in thenasal passages of a patient comprising: administering rhCClO to thepatient.
 2. The method of claim 1 wherein the amount of rhCClOadministered is between about 1.5 micrograms and about 1.5 milligramsper day.
 3. The method of claim 1 wherein the amount of rhCClOadministered is less than about 1.1 milligrams per day.
 4. The method ofclaim 1 wherein the amount of rhCClO that is administered is about 0.75micrograms to about 650 micrograms per day.
 5. The method of claim 1wherein the amount of rhCClO that is administered is about 0.5micrograms to about 370 micrograms per day.
 6. The method of claim 1wherein the administration of rhCClO is repeated about three times perday.
 7. The method of claim 1 wherein the administration of rhCClO isrepeated about two days.
 8. The method of claim 1 wherein rhCClO isadministered to the nasal passages by instillation, lavage, swabapplicator, or spray.
 9. The method of claim 1 wherein rhCClO isadministered in combination with an antibiotic, an anti-histamine, adecongestant, a mucolytic, an analgesic, a local-acting vasoconstrictor,a leukotriene receptor antagonist, a steroid, a nasal excipient, or anycombination thereof.
 10. A safe and well-tolerated method of preventingor slowing growth or regrowth of nasal polyps in a patient comprising:administering rhCClO to the patient.
 11. The method of claim 10 whereinrhCC 10 between about 1.5 micrograms and 1.5 milligrams is administeredeach day for at least 2 days.
 12. The method of claim 10 wherein theamount of rhCClO that is administered is 1.1 milligrams per day or less.13. The method of claim 10 wherein the administration of rhCClO isrepeated about three times per day.
 14. The method of claim 10 whereinrhCClO is administered to the nasal passages by instillation, lavage,swab applicator, or spray.
 15. The method of claim 10 wherein rhCClO isadministered in combination with an antibiotic, an anti-histamine, adecongestant, a mucolytic, an analgesic, a local-acting vasoconstrictor,a leukotriene receptor antagonist, a steroid, a nasal excipient, or anycombination thereof.
 16. A safe and well-tolerated method of treating orpreventing chronic or recurrent bacterial sinus infection in a patientcomprising: administering rhCClO to the patient.
 17. The method of claim16 wherein rhCClO between about 1.5 micrograms and 1.5 milligrams isadministered each day for at least 2 days.
 18. The method of claim 16wherein the amount of rhCC 10 that is administered is 1.1 milligrams perday or less.
 19. The method of claim 16 wherein the administration ofrhCClO is repeated about three times per day.
 20. The method of claim 16wherein rhCClO is administered to the nasal passages by instillation,lavage, swab applicator, or spray.
 21. The method of claim 16 whereinrhCClO is administered in combination with an antibiotic, ananti-histamine, a decongestant, a mucolytic, an analgesic, alocal-acting vasoconstrictor, a leukotriene receptor antagonist, asteroid, a nasal excipient, or any combination thereof.
 22. A safe andwell-tolerated method of treating sinus pain in a patient comprising:administering rhCClO to the patient.
 23. The method of claim 22 whereinrhCClO between about 1.5 micrograms and 1.5 milligrams is administeredeach day for at least 2 days.
 24. The method of claim 22 wherein theamount of rhCClO that is administered is 1.1 milligrams per day or less.25. The method of claim 22 wherein the administration of rhCClO isrepeated about three times per day.
 26. The method of claim 22 whereinrhCClO is administered to the nasal passages by instillation, lavage,swab applicator, or spray.
 27. The method of claim 22 wherein rhCC 10 isadministered in combination with an antibiotic, an anti-histamine, adecongestant, a mucolytic, an analgesic, a local-acting vasoconstrictor,a leukotriene receptor antagonist, a steroid, a nasal excipient, or anycombination thereof.
 28. A pharmaceutical composition for intranasaladministration of rhCClO comprising a formulation comprising rhCClO incombination with a nasal excipient.
 29. The pharmaceutical compositionof claim 28 comprising rhCClO at a concentration no greater than 2milligrams/milliliter.
 30. The pharmaceutical composition of claim 28comprising rhCClO at a concentration of 250 micrograms per milliliter in0.65% sodium chloride, disodium phosphate, phenylcarbinol, monosodiumphosphate, and benzalkonium chloride at a pH of 4.0-8.0.
 31. (canceled)32. A drug-device combination composition comprising the pharmaceuticalcomposition of claim 30 and a squeeze spray bottle, pump action spraydevice, metered dose nasal actuator, syringe-type instillation device,nasal swab applicator, or “Neti pot” lavage device that enables thetopical application of rhCClO to the surfaces of the nasal passages. 33.The method of claim 1 wherein rhCClO is administered in a pharmaceuticalcomposition comprising a formulation comprising rhCClO in combinationwith a nasal excipient wherein rhCC10 is present in a concentration nogreater than 2 milligrams/milliliter in a dosage of 20-50 micrograms pernostril up to four times per day.
 34. The method of claim 10 whereinrhCClO is administered in a pharmaceutical composition comprising aformulation comprising rhCClO in combination with a nasal excipientwherein rhCC10 is present in a concentration no greater than 2milligrams/milliliter in a dosage of 20-50 micrograms per nostril up tofour times per day.
 35. The method of claim 16 wherein rhCClO isadministered in a pharmaceutical composition comprising a formulationcomprising rhCClO in combination with a nasal excipient wherein rhCC10is present in a concentration no greater than 2 milligrams/milliliter ina dosage of 20-50 micrograms per nostril up to four times per day. 36.The method of claim 20 wherein rhCClO is administered in apharmaceutical composition comprising a formulation comprising rhCClO incombination with a nasal excipient wherein rhCC10 is present in aconcentration no greater than 2 milligrams/milliliter in a dosage of20-50 micrograms per nostril up to four times per day.